Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis

被引：28
|
作者：
Emmett, Mark R.
Kazazic, Sasa
Marshall, Alan G.
Chen, Wei
Shi, Stone D. -H.
Bolanos, Ben
Greig, Michael J.
机构：
[1] Florida State Univ, Natl High Magnet Field Lab, Ion Cyclotron Resonance Program, Tallahassee, FL 32310 USA
[2] Florida State Univ, Dept Chem & Biochem, Tallahassee, FL 32306 USA
[3] Pfizer Inc, Global Res & Dev, La Jolla Labs, San Diego, CA 92121 USA
来源：
ANALYTICAL CHEMISTRY | 2006年 / 78卷 / 19期
关键词：
PROTEIN-PROTEIN INTERACTIONS; CHEMICAL CROSS-LINKING; IDENTIFICATION; PEPTIDES; SURFACES;
D O I：
10.1021/ac060693n
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
The single biggest problem with solution-phase H/D exchange as a mass spectrometric probe of surface exposure in a protein (or protein complex) is back-exchange of H for D after the initial H/D exchange has been quenched. Back-exchange results in loss of pertinent data and also greatly hampers data analysis. Previously, very fast, cold (0-4 degrees C) HPLC was performed to help reduce back-exchange, but calculated back-exchange still averages similar to 30%. In this report, supercritical fluid chromatography replaces HPLC as the desalting/separation technique prior to mass analysis, providing a dramatic reduction in back-exchange compared to the fast, cold HPLC methods.
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页码：7058 / 7060
页数：3
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